title: Role of CcpA in polyhydroxybutyrate biosynthesis in a newly isolated Bacillus sp. MA3.3.
abstract: The ccpA gene was inactivated in the polyhydroxybutyrate (PHB)-producing strain Bacillus sp. MA3.3 in order to reduce glucose catabolite repression over pentoses and develop improved bacterial strains for the production of PHB from lignocellulosic hydrolysates. Mutant Bacillus sp. MSL7 ΔCcpA are unable to grow on glucose and ammonia as sole carbon and nitrogen sources, respectively. Supplementation of glutamate as the nitrogen source or the substitution of the carbon source by xylose allowed the mutant to partially recover its growth performance. RT-PCR showed that CcpA stimulates the expression of the operon (gltAB),responsible for ammonia assimilation via glutamate in Bacillus sp. MA3.3. Moreover, it was demonstrated that the supplementation of xylose or glutamate was capable of stimulating gltAB operon expression independently of CcpA. In PHB production experiments in mineral media, it has been observed that the glucose catabolite repression over the pentoses was partially released in MSL7. Although the carbohydrate consumption is faster in the ccpA mutant, the biomass and PHB biosynthesis are lower, even with supplementation of glutamate. This is attributed to an increase of acetyl-CoA flux towards the tricarboxylic acid cycle observed in the mutant.
authors.affiliation: Departamento de Microbiologia, Instituto de Ciências Biomédicas, Universidade de São Paulo, São Paulo, Brazil.
authors.forename: M S L
authors.initials: MS
authors.lastname: Lopes
authors.forename: N
authors.initials: N
authors.lastname: Steinert
authors.forename: J D
authors.initials: JD
authors.lastname: Rojas
authors.forename: W
authors.initials: W
authors.lastname: Hillen
authors.forename: J G C
authors.initials: JG
authors.lastname: Gomez
authors.forename: L F
authors.initials: LF
authors.lastname: Silva
mesh_terms: D000641:Ammonia / Q000378:metabolism; D001407:Bacillus / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification / Q000378:metabolism*; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D020869:Gene Expression Profiling; D055786:Gene Knockout Techniques; D005947:Glucose / Q000378:metabolism; D018698:Glutamic Acid / Q000378:metabolism; D006885:Hydroxybutyrates / Q000378:metabolism*; D011091:Polyesters / Q000378:metabolism*; D014994:Xylose / Q000378:metabolism; D000094122:Polyhydroxybutyrates
chemical_list: D000641:Ammonia; D001426:Bacterial Proteins; D005947:Glucose; D018698:Glutamic Acid; D006885:Hydroxybutyrates; D011091:Polyesters; D014994:Xylose; C003182:poly-beta-hydroxybutyrate; D000094122:Polyhydroxybutyrates